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目的探讨骨髓移植后供体骨髓来源干细胞(bone marrow derived stem cells,BMDSCs)是否能够在受体肠上皮分化为具有正常超微结构和功能的肠上皮细胞。方法使用Percoll液分离GFP转基因小鼠骨髓有核细胞,经尾静脉移植后3个月取材。标本经戊二醛固定,饿酸后固定,常规电镜超薄切片。采用包埋后染色免疫电镜技术以GFP抗体识别供体细胞,5nm胶体金标记二抗识别一抗后,进行电镜染色和观察。结果透射电镜下可见电子密度高的单个或成簇分布的球形胶体金颗粒散布于肠上皮细胞内,阴性对照组未见胶体金颗粒。阳性细胞主要为杯状细胞和吸收细胞,且阳性细胞同其他上皮细胞之间存在有细胞连接。结论通过免疫电镜技术证实,移植后的供体骨髓来源细胞可以在受体小肠组织中分化为具有正常功能的肠上皮细胞。
Objective To investigate whether bone marrow derived stem cells (BMDSCs) can differentiate into intestinal epithelial cells with normal ultrastructure and function in recipient intestine after bone marrow transplantation. Methods The bone marrow cells of GFP transgenic mice were isolated by Percoll fluid and drawn at 3 months after tail vein transplantation. Specimens were fixed by glutaraldehyde, hungry after fixation, conventional ultra-thin sections of electron microscopy. After embedding, the immunofluorescence microscopy was used to identify the donor cells with GFP antibody. After the 5nm colloidal gold labeled secondary antibody was used to identify the primary antibody, the cells were stained with electron microscopy and observed. Results Single or clustered spherical colloidal gold particles with high electron density were found in the intestinal epithelial cells under transmission electron microscope. No colloidal gold particles were found in the negative control group. Positive cells are goblet cells and absorptive cells, and positive cells with other epithelial cells there is a cell connection. Conclusions Immuno-electron microscopy confirmed that the donor-derived bone marrow-derived cells could differentiate into normal intestinal epithelial cells in the recipient small intestine.